| Title: |
A standardised methodology for the extraction and quantification of cell-free DNA in cerebrospinal fluid and application to evaluation of Alzheimer's disease and brain cancers. |
| Authors: |
Takousis, Petros; Devonshire, Alison S; Huggett, Jim F; Perneczky, Robert; Redshaw, Nicholas; von Baumgarten, Louisa; Whale, Alexandra S; Jones, Gerwyn M; Fernandez-Gonzalez, Ana; Martin, Jan; Foy, Carole A; Alexopoulos, Panagiotis |
| Source: |
New biotechnology 72, 97 - 106 (2022). doi:10.1016/j.nbt.2022.10.001 |
| Publisher Information: |
Elsevier |
| Publication Year: |
2022 |
| Subject Terms: |
info:eu-repo/classification/ddc/540; Humans; Cell-Free Nucleic Acids; Alzheimer Disease: diagnosis; Biomarkers; Brain Neoplasms; Dementia; biomarker; brain tumour; diagnosis; genetics; metastasis |
| Subject Geographic: |
DE |
| Description: |
Cerebrospinal fluid (CSF) is a source of diagnostic biomarkers for a range of neurological conditions. Cell-free DNA (cfDNA) is detected in CSF and differences in the concentration of cell-free mitochondrial DNA have been reported in studies of neurodegenerative disorders including Alzheimer's disease (AD). However, the influence of pre-analytical steps has not been investigated for cfDNA in CSF and there is no standardized approach for quantification of total cfDNA (copies of nuclear genome or mitochondria-derived gene targets). In this study, the suitability of four extraction methods was evaluated: QIAamp Circulating Nucleic Acid (Qiagen), Quick-cfDNA Serum & Plasma (Zymo), NucleoSnap® DNA Plasma (Macherey-Nagel) and Plasma/Serum Circulating DNA Purification Mini (Norgen) kits, for cfDNA extraction from CSF of controls and AD dementia patients, utilising a spike-in control for extraction efficiency and fragment size. One of the optimal extraction methods was applied to a comparison of cfDNA concentrations in CSF from control subjects, AD dementia and primary and secondary brain tumour patients. Extraction efficiency based on spike-in recovery was similar in all three groups whilst both endogenous mitochondrial and nucleus-derived cfDNA was significantly higher in CSF from cancer patients compared to control and AD groups, which typically contained < 100 genome copies/mL. This study shows that it is feasible to measure low concentration nuclear and mitochondrial gene targets in CSF and that normalization of extraction yield can help control pre-analytical variability influencing biomarker measurements. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| ISSN: |
1871-6784; 1876-4347 |
| Relation: |
info:eu-repo/semantics/altIdentifier/issn/1871-6784; info:eu-repo/semantics/altIdentifier/pmid/pmid:36202346; info:eu-repo/semantics/altIdentifier/issn/1876-4347; https://pub.dzne.de/record/165317 |
| Availability: |
https://pub.dzne.de/record/165317; https://pub.dzne.de/search?p=id:%22DZNE-2022-01595%22 |
| Rights: |
info:eu-repo/semantics/openAccess |
| Accession Number: |
edsbas.3767165B |
| Database: |
BASE |